CN106011132A - Method for extracting corn or radish genome DNA with high flux - Google Patents
Method for extracting corn or radish genome DNA with high flux Download PDFInfo
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- CN106011132A CN106011132A CN201610672468.4A CN201610672468A CN106011132A CN 106011132 A CN106011132 A CN 106011132A CN 201610672468 A CN201610672468 A CN 201610672468A CN 106011132 A CN106011132 A CN 106011132A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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Abstract
The invention discloses a method for extracting corn or radish genome DNA with high flux. The method is characterized in that a leaves sample of corn or radish to-be-detected can be obtained with batches, then alkali lye is added, cell wall and cell membrane of the plant leaves can be destroyed under high temperature condition, DNA is released, an acidic neutralization extract is added so that the sample is suitable for PCR amplification, and then an amplification reaction is carried out to realize detection of target molecule. The method has the advantages that operation is simple, the method is benefit for manual operation, is suitable for operation by a miniature laboratory, and the processing efficiency is high.
Description
Technical field
Agriculture plant breeding DNA extraction technical field of the present invention, is specifically related to a kind of high flux and extracts Semen Maydis or radish gene group
The method of DNA.
Background technology
Crop breeding, also known as breed improvement, is by creatingHereditary variation, improvement inherited character, to cultivate excellent animals and plants
New varieties so that it is reach the technology of high yield, stable yields, high-quality, excellent results.To developing animal husbandry and plant husbandry has particularly significant
Meaning.During crop breeding, use is generally individually needed to arrivePolymerase chain reactionTechnology realizes molecular marker,Polymerase chain Formula is reacted(PCR) it is a kind of for amplifying the specific DNA fragmentation of amplificationProtocols in Molecular Biology, it is considered as in vitro
Special DNA replication dna, the maximum feature of PCR, is to be significantly increased by the DNA of trace.
Semen Maydis based on polymerase chain reaction, Radix Raphani molecular mark need to identify in big colony and contain
There are individual plant or the strain of objective trait (degeneration-resistant, high-quality).In molecular breeding is put into practice, one of method is that research worker is by breeding group
Body is pressed simple grain and is germinateed in seedbed, and seedling completes the Molecular Identification to objective trait before moving into field planting.The two of method are
Directly seed is carried out Molecular Detection.While it is true, research worker would generally be at a breeding generation to multiple breeding populations
Thousands of parts of individual plants carry out the Molecular Identification of objective trait.Therefore, quick obtaining number becomes with thousand parts of DNA that can be used for PCR amplification
One of factor of restriction molecule labelling efficiency of selection.
At present, the method extracting DNA of plants conventional has CTAB(cetyl trimethylammonium bromide) method, SDS(dodecyl
Sodium sulfate) method such as method, DNA commercial reagents box.
CTAB method and SDS method major defect are that process is complicated, loaded down with trivial details time-consumingly.Extract Radix Raphani and maize dna needs 8 to 10
Step, every part of sample needs to be fully ground in liquefied ammonia, has cross-contamination unavoidably, need subsequently during grinding between sample
Long-time concussion, is centrifuged repeatedly, extracts and washs, and is finally dried and dissolves.1 day multipotency of 1 skilled technician extracts 30 parts
Sample, this is far from the needs meeting molecular mark practice.
DNA commercial reagents box major defect is expensive, and use cost is high.Molecule assisted Selection is applied to the master of breeding
Want advantage can improve breeding efficiency, reduce breeding cost.But, utilize commercial reagents box to extract DNA and carry out Molecular Detection by bright
The aobvious cost improving molecule assisted Selection, thus add breeding cost.
Summary of the invention
For above-mentioned the deficiencies in the prior art, the technical problem to be solved is: how to provide a kind of for jade
Rice, Radix Raphani DNA, simple to operate, beneficially manual operation, with low cost, and the high high flux for the treatment of effeciency extracts Semen Maydis or Radix Raphani base
Because of the method organizing DNA.
In order to solve above-mentioned technical problem, present invention employs following technical scheme:
A kind of high flux extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that first obtain jade to be detected in bulk
Rice or the leaf sample of Radix Raphani, be then respectively adding alkali liquor, destroy plant leaf blade cell wall and cell membrane under the high temperature conditions,
Being discharged by DNA, the neutralising extract being subsequently adding acidity is allowed to be suitable for PCR amplification, then realizes mesh after carrying out amplified reaction
Mark Molecular Detection.
This method, specifically includes following steps:
A, the seed simple grain of batch Semen Maydis to be detected or Radix Raphani is cultivated grow blade；
B, it is sampled according to the 30-60 square millimeter each Semen Maydis to cultivating or radish plant blade, and puts into process orifice plate
In each aperture；
C, preparation buffer A and buffer B, buffer A is 0.1mol/L NaOH and 2% Tween20 equal-volume is mixed to get,
Buffer B: 0.1mol/L Tris-HCl and 0.002 mol/L EDTA equal-volume are mixed to get；To each aperture of process orifice plate
After middle addition 500ul buffer A, process orifice plate is heated a period of time；
D, again addition 500ul buffer B in each aperture of process orifice plate, shake a period of time so that liquid is sufficiently mixed, and releases
Release DNA；
E, PCR reaction system is distributed in each aperture of reaction orifice plate, process orifice plate after concussion in corresponding removing step d
During liquid enters into each aperture of reaction orifice plate in each aperture, carry out amplified reaction, then realize objective trait Molecular Detection.
The cardinal principle of this method is that alkalescence, the condition of high temperature can destroy plant leaf blade cell wall and cell membrane, thus will
DNA discharges, and then is allowed to be suitable for PCR amplification with acid neutralising extract.Concentration Tween20 within 2% can crack carefully
After birth released dna, again will not suppression PCR amplified reaction.Therefore this method need not be ground sample and pretreatment, for little
The current demand of type laboratory, by the nucleic acid extraction of high flux low cost, under conditions of being not required to mechanical hand, can realize full
The Molecular Detection of the high flux objective trait of foot breeding demand.
As optimization, in a step, the seed simple grain of batch Semen Maydis to be detected or Radix Raphani is respectively put into breeding hole
In each aperture of plate, cultivate in illumination box and grow blade；Breeding orifice plate preferably employs 96 orifice plates.So can ensure that and criticize
The efficiency that amount processes.
In b step, it is preferred to use plant leaf blade sampler is sampled, and process orifice plate preferably employs 96 hole PCR plate
Guaranteed efficiency.
In step c, buffer is allocated and can be used following methods:
(1) 100mM NaOH: being dissolved in by 0.2 g NaOH in double pure water, constant volume is 50 ml；
(2) 2% Tween20: take 5 g 20% Tween20 and be dissolved in double pure water of 50 ml；
(3) 100mM Tris-HCl: taking 0.605 g Tris-HCl and be dissolved in double pure water, regulate PH=8, constant volume is 50ml；
(4) 2mM EDTA: taking 0.037 g EDTA and be dissolved in double pure water, regulate PH=8, constant volume is 50ml；
(5) buffer A: 0.1mol/L NaOH and the mixing of 2% Tween20 equal-volume, now with the current；
(6) buffer B: 0.1mol/L Tris-HCl and the mixing of 0.002 mol/L EDTA equal-volume.
Use said method, convenient enforcement to allocate, be especially advantageous in small test room environmental implementing preparation.
In step c, it is preferred to use the 12 passage liquid-transfering guns of range 1000ul add buffer A, to facilitate operation and to improve
Efficiency；Wherein heating purpose is to utilize alkalescence, the condition destruction plant leaf blade cell wall of high temperature and cell membrane, thus is released by DNA
Release, it is preferred to use in thermal cycler, 95 DEG C of heating 10min, can just reach this effect and be unlikely to again to destroy DNA.
In Step d, use and on earthquake device, shake 5min, make buffer A and buffer B be sufficiently mixed；To ensure mixing
Fully, DNA can fully be discharged in solution.
In step e, reaction orifice plate preferably employs 96 hole PCR plate guaranteed efficiencies；Liquid pipettes and preferably employs range
The 12 passage liquid-transfering guns of 1000ul, to facilitate operation and to improve efficiency；0.1% can also be added further in PCR reaction system
Bovine serum albumin (BSA) and 1% polyvinylpyrrolidone (PVP) can effectively reduce the effect of inhibitive factor, improve detection effect
Really.
This method has the following characteristics that 1, most important advantage is high flux and low cost.Utilize the method extract DNA, 1
Individual research worker can process more than 2000 sample for 1 day, and the cost extracting 1 DNA is approximately 0.05 dollar.Than CTAB and SDS method
Save time, more cheap than test kit.2, the blade amount needed is few.It is about 30-60mm2Enough 100 PCR of DNA that extract of blade expand
Increase reaction.3, while having above-mentioned advantage, the DNA that the method is extracted can stably preserve 1 month, at-20 DEG C at 4 DEG C
Stably preserve 3 months.4, extract DNA by the method, carry out pcr amplification reaction in this, as template, be not required to Robot actions
In the case of, it is possible to realize sequencing and the standardization of Molecular Detection.Therefore, application the method extraction DNA is very suitable for little
Type labs molecular marker assisted selection works.
In sum, the present invention has simple to operate, beneficially manual operation, and applicable small-size laboratory is carried out, with low cost,
And treatment effeciency advantages of higher.
Accompanying drawing explanation
Fig. 1 is to take Radix Raphani DNA extraction sample liquid during Radix Raphani verification experimental verification to carry out agarose gel electrophoresis detection, different turnip leaves
The result schematic diagram of the extracting solution detection of taken amount.
Fig. 2 is to take Radix Raphani DNA extraction sample liquid during Radix Raphani verification experimental verification to carry out agarose gel electrophoresis detection, and sampling amount is 5mg
Radix Raphani DNA extraction liquid agarose gel electrophoresis figure.
Radix Raphani DNA extraction liquid PCR primer detection figure when Fig. 3 is Radix Raphani verification experimental verification.
Fig. 4 is to take maize dna extraction sample liquid during corn trials checking to carry out agarose gel electrophoresis detection, different leaf of Semen Maydis
The result schematic diagram of the extracting solution detection of sheet taken amount.
Fig. 5 is to take maize dna extraction sample liquid during corn trials checking to carry out agarose gel electrophoresis detection, and sampling amount is
The maize dna extracting solution agarose gel electrophoresis figure of 10mg.
Maize dna extracting solution sepharose electrophoresis detection figure when Fig. 6 is corn trials checking.
Detailed description of the invention
The present invention is described in further detail with optimum embodiment below in conjunction with the accompanying drawings.
Optimum embodiment: a kind of high flux extracts Semen Maydis or the method for radish gene group DNA, specifically includes following steps:
A, the seed simple grain of batch Semen Maydis to be detected or Radix Raphani is cultivated grow blade；
B, it is sampled according to the 30-60 square millimeter each Semen Maydis to cultivating or radish plant blade, and puts into process orifice plate
In each aperture；
C, preparation buffer A and buffer B, buffer A is 0.1mol/L NaOH and 2% Tween20 equal-volume is mixed to get,
Buffer B: 0.1mol/L Tris-HCl and 0.002 mol/L EDTA equal-volume are mixed to get；To each aperture of process orifice plate
After middle addition 500ul buffer A, process orifice plate is heated a period of time；
D, again addition 500ul buffer B in each aperture of process orifice plate, shake a period of time so that liquid is sufficiently mixed, and releases
Release DNA；
E, PCR reaction system is distributed in each aperture of reaction orifice plate, process orifice plate after concussion in corresponding removing step d
During liquid enters into each aperture of reaction orifice plate in each aperture, carry out amplified reaction, then realize objective trait Molecular Detection.
In present embodiment, in a step, the seed simple grain of batch Semen Maydis to be detected or Radix Raphani is respectively put into breeding
With in each aperture of orifice plate, cultivate in illumination box and grow blade；Breeding orifice plate uses 96 orifice plates.In b step, employing is planted
Thing blade sampler is sampled, and process orifice plate uses 96 hole PCR plate.In step c, buffer allotment can use below
Method:
(1) 100mM NaOH: being dissolved in by 0.2 g NaOH in double pure water, constant volume is 50 ml；
(2) 2% Tween20: take 5 g 20% Tween20 and be dissolved in double pure water of 50 ml；
(3) 100mM Tris-HCl: taking 0.605 g Tris-HCl and be dissolved in double pure water, regulate PH=8, constant volume is 50ml；
(4) 2mM EDTA: taking 0.037 g EDTA and be dissolved in double pure water, regulate PH=8, constant volume is 50ml；
(5) buffer A: 0.1mol/L NaOH and the mixing of 2% Tween20 equal-volume, now with the current；
(6) buffer B: 0.1mol/L Tris-HCl and the mixing of 0.002 mol/L EDTA equal-volume.
In present embodiment step c, the 12 passage liquid-transfering guns of range 1000ul are used to add buffer A.Use thermal cycle
95 DEG C of instrument heating 10min.In Step d, use and on earthquake device, shake 5min, make buffer A and buffer B be sufficiently mixed.e
In step, reaction orifice plate uses 96 hole PCR plate, and liquid pipettes the 12 passage liquid-transfering guns preferably employing range 1000ul.Simultaneously
0.1% bovine serum albumin and 1% polyvinylpyrrolidone is added in PCR reaction system.
Below, it is respectively directed to radish seed and corn seed, based on above-mentioned detailed description of the invention operating procedure, uses
Tests below verifies effect of the present invention.
One, Radix Raphani verification experimental verification.
The extraction of Radix Raphani DNA and detection.
1, radish seed is put in incubator germinate, blade sampling gradient be preset as 2 mg, 5 mg, 10 mg, 15 mg,
20 mg、25 mg、30 mg、35 mg、40 mg、50 mg。
2, DNA extraction is completed by the operating procedure in detailed description of the invention.
3, DNA detection: take Radix Raphani DNA extraction sample liquid and carry out agarose gel electrophoresis detection.It is illustrated in figure 1 different Folium Raphani
The extracting solution detection figure of sheet taken amount, in figure: M represents DNAmarker；；1～10 loading wells are all the different sampling amounts of same volume
DNA extraction liquid, be followed successively by from left to right 50 mg, 40 mg, 35 mg, 30 mg, 25 mg, 20 mg, 15 mg, 10 mg, 5
Mg, the DNA extraction liquid of turnip leaves taken amount of 2 mg.As shown in Figure 1: blade sampling amount does not the most affect the dense of DNA in extracting solution
Degree.
Fig. 2 be sampling amount be the Radix Raphani DNA extraction liquid agarose gel electrophoresis figure of 5mg, in figure: M represents marker；1～13
Loading wells is followed successively by extracting solution 6.0 μ l, 5.75 μ l, 5.5 μ l, 5.25 μ l, 5.0 μ l, 4.75 μ l, 4.5 μ l, 4.25 μ
l、4.0 μl、3.75 μl、3.50 μl、3.25 μl、3.0 μl.As seen from the figure: utilize this method can extract Radix Raphani DNA, extract
In liquid, the concentration containing the DNA, DNA discharged increases along with the extracting liquid volume of detection and improves.
4, the pcr amplification reaction with this DNA as template.
PCR reaction system (25 μ l): 12.5 μ l 2 × Taq Master Mix, ITSR upper and lower primer (10 nmol/
Ml) each 1 μ l, DNA extraction liquid 3 μ l, Nase Free Water 7.5 μ l.
PCR response procedures:
5, amplified reaction electrophoresis detection is carried out, as it is shown on figure 3, be Radix Raphani DNA extraction liquid PCR primer detection figure, in figure: M represents
marker；1-5 is followed successively by blade sampling amount 5 mg； 10 mg；15 mg ；20 mg； 25 mg ；The primer is ITSR,
The band of its amplification is it is contemplated that 750bp.As shown in Figure 3: the DNA extraction liquid of all taken amounts can amplify expection product, PCR expands
Agarose gel electrophoresis effect and the blade sampling amount of volume increase thing number unrelated.The DNA extraction liquid that the method is obtained as
Pcr template, it is possible to amplify expection band, it was demonstrated that the method feasibility in Radix Raphani Molecular Detection.
Two, corn trials checking.
The extraction of maize dna and detection.
1, corn seed is put in incubator germinate, blade sampling gradient be preset as 2 mg, 5 mg, 10 mg, 15 mg,
20 mg、25 mg、30 mg、35 mg、40 mg、50 mg。
2, DNA extraction is completed by the operating procedure in detailed description of the invention.
3, DNA detection: take maize dna extraction sample liquid and carry out agarose gel electrophoresis detection.It is not that result sees Fig. 4, Fig. 4
DNA agarose gel electrophoresis figure is extracted with blade taken amount.In figure: M represents DNA marker；1～10 loading wells are all identical
The DNA extraction liquid of volume difference sampling amount.Be followed successively by from left to right 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30
Mg, 35 mg, 40 mg, the DNA extraction liquid of different maize leaf taken amounts of 50 mg.As shown in Figure 4: all blade taken amounts DNA carry
Take liquid and all can detect DNA.And can be seen that the extracting solution DNA concentration of 10 mg blades is the highest.Fig. 5 be sampling amount be 10mg's
Maize dna extracting solution agarose gel electrophoresis figure.In figure: M represents DNAmarker；1-6 is followed successively by extracting solution 5 μ l, 4.5 μ
L, 4 μ l, 3.5 μ l, 3 μ l, 2.5 μ l, 7 bromophenol blues.As shown in Figure 5: pipette 4 μ l DNA extraction liquid and carry out Detection results
Best.
4, the pcr amplification reaction with this DNA as template.
PCR reaction system (25 μ l): 12.5 μ l 2 × Taq Master Mix, ITSR upper and lower primer (10 nmol/
Ml) each 0.5 μ l, DNA are 1 μ l, Nase Free Water 10.5 μ l.
PCR response procedures:
5, amplified reaction electrophoresis detection.Fig. 6 is maize dna extracting solution sepharose electrophoresis detection figure；In figure: M represents
DNAmarker；1～10 be followed successively by blade sampling amount 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg,
40 mg、50 mg.The primer is ITSR, and it amplifies band it is contemplated that 750bp.As shown in Figure 6: the DNA of all taken amounts carries
Taking liquid and can amplify expection product, sample taken amount is 2 mg, 5 mg, 10 mg also can amplify product, but be not reaching to pre-
Phase effect.Can be seen that the PCR amplification of Semen Maydis is relevant to blade sampling amount, and be gradually increased to a certain extent at blade sampling amount
Time on PCR reaction impact little.Semen Maydis extracts the leaf quality of DNA preferably between 10 mg～30 mg.
Test Summary: to sum up, utilizes the method can extract DNA from Semen Maydis, Radix Raphani, and the DNA extracted disclosure satisfy that PCR
The requirement of amplified reaction, small-size laboratory uses the method, it is possible to realize high flux and low cost.
Claims (10)
1. a high flux extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that first obtain to be detected in bulk
Semen Maydis or the leaf sample of Radix Raphani, be then respectively adding alkali liquor, destroys plant leaf blade cell wall and cell under the high temperature conditions
Film, discharges DNA, and the neutralising extract being subsequently adding acidity is allowed to be suitable for PCR amplification, then carries out realization after amplified reaction
Target molecule detects.
2. high flux as claimed in claim 1 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that specifically include
Following steps:
A, the seed simple grain of batch Semen Maydis to be detected or Radix Raphani is cultivated grow blade；
B, it is sampled according to the 30-60 square millimeter each Semen Maydis to cultivating or radish plant blade, and puts into process orifice plate
In each aperture；
C, preparation buffer A and buffer B, buffer A is 0.1mol/L NaOH and 2% Tween20 equal-volume is mixed to get,
Buffer B: 0.1mol/L Tris-HCl and 0.002 mol/L EDTA equal-volume are mixed to get；To each aperture of process orifice plate
After middle addition 500ul buffer A, process orifice plate is heated a period of time；
D, again addition 500ul buffer B in each aperture of process orifice plate, shake a period of time so that liquid is sufficiently mixed, and releases
Release DNA；
E, PCR reaction system is distributed in each aperture of reaction orifice plate, process orifice plate after concussion in corresponding removing step d
During liquid enters into each aperture of reaction orifice plate in each aperture, carry out amplified reaction, then realize objective trait Molecular Detection.
3. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that in a step,
The seed simple grain of batch Semen Maydis to be detected or Radix Raphani is respectively put in each aperture of breeding orifice plate, in illumination box
Cultivation grows blade；Breeding orifice plate uses 96 orifice plates.
4. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that in b step,
Use plant leaf blade sampler to be sampled, and process orifice plate uses 96 hole PCR plate.
5. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that in step c,
Buffer is allocated and can be used following methods:
(1) 100mM NaOH: being dissolved in by 0.2 g NaOH in double pure water, constant volume is 50 ml；
(2) 2% Tween20: take 5 g 20% Tween20 and be dissolved in double pure water of 50 ml；
(3) 100mM Tris-HCl: taking 0.605 g Tris-HCl and be dissolved in double pure water, regulate PH=8, constant volume is 50ml；
(4) 2mM EDTA: taking 0.037 g EDTA and be dissolved in double pure water, regulate PH=8, constant volume is 50ml；
(5) buffer A: 0.1mol/L NaOH and the mixing of 2% Tween20 equal-volume, now with the current；
(6) buffer B: 0.1mol/L Tris-HCl and the mixing of 0.002 mol/L EDTA equal-volume.
6. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that in step c,
The 12 passage liquid-transfering guns using range 1000ul add buffer A.
7. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that in step c,
Use thermal cycler 95 DEG C heating 10min.
8. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that in Step d,
Use and on earthquake device, shake 5min, make buffer A and buffer B be sufficiently mixed.
9. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that in step e,
Reaction orifice plate uses 96 hole PCR plate, and liquid pipettes the 12 passage liquid-transfering guns preferably employing range 1000ul.
10. high flux as claimed in claim 2 extracts Semen Maydis or the method for radish gene group DNA, it is characterised in that step e
In, PCR reaction system adds 0.1% bovine serum albumin and 1% polyvinylpyrrolidone.
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| CN201610672468.4A CN106011132A (en) | 2016-08-16 | 2016-08-16 | Method for extracting corn or radish genome DNA with high flux |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107201363A (en) * | 2017-07-24 | 2017-09-26 | 江苏大学 | A kind of simplicity of DNA of plants, rapid extracting method |
| CN110184266A (en) * | 2019-06-03 | 2019-08-30 | 中国农业科学院植物保护研究所 | Citrus leaf DNA rapid extracting method and its application in Citrus Huanglongbing pathogen detection |
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| CN1308130A (en) * | 2000-12-26 | 2001-08-15 | 李永武 | Method of extracting nucleic and from microbe cell |
| CN103436524A (en) * | 2013-06-17 | 2013-12-11 | 华南农业大学 | Method for batch extraction of rice endosperm DNA |
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- 2016-08-16 CN CN201610672468.4A patent/CN106011132A/en not_active Withdrawn
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1308130A (en) * | 2000-12-26 | 2001-08-15 | 李永武 | Method of extracting nucleic and from microbe cell |
| CN103436524A (en) * | 2013-06-17 | 2013-12-11 | 华南农业大学 | Method for batch extraction of rice endosperm DNA |
Non-Patent Citations (1)
| Title |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107201363A (en) * | 2017-07-24 | 2017-09-26 | 江苏大学 | A kind of simplicity of DNA of plants, rapid extracting method |
| CN110184266A (en) * | 2019-06-03 | 2019-08-30 | 中国农业科学院植物保护研究所 | Citrus leaf DNA rapid extracting method and its application in Citrus Huanglongbing pathogen detection |
| CN110184266B (en) * | 2019-06-03 | 2021-06-04 | 中国农业科学院植物保护研究所 | Rapid extraction method of citrus leaf DNA and application of rapid extraction method in citrus yellow shoot detection |
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